Fas activation induces renal tubular epithelial cell beta 8 integrin expression and function in the absence of apoptosis

Cell fate following Fas (CD95) ligand or agonistic anti-Fas antibody stimulation is determined by multiple factors, including Fas expression level, microdomain localization, and modulating cytokines. Highly expressed Fas clusters and activates a canonical apoptosis signaling pathway. In less susceptible cells, Fas transduces apoptosis-independent signals, which are not well defined, but have been linked to inflammation, angiogenesis, and fibrosis. To identify apoptosis-independent Fas pathways, cultured renal tubular epithelial cells were stimulated with agonistic anti-Fas antibodies under conditions that did not cause cell death. Analysis of filter cDNA microarrays revealed beta(8) integrin subunit mRNA induction in Fas-stimulated cells. beta(8) integrin mRNA expression increased within 3-6 h of Fas ligation due to enhanced mRNA stabilization, and mRNA increases were sustained for 48-72 h. Expression of plasma membrane beta(8) integrin, as well as its heterodimer partner alpha(v), was increased by Fas activation with a similar kinetic pattern. Fas-induced alpha(v)beta(8) expression correlated with increased migration to vitronectin, the ligand for alpha(v)beta(8). Results from studies with function-blocking antibodies against other alpha(v)beta integrins or suppression of beta(8) integrin expression by RNA interference demonstrated that induced beta(8) integrin expression mediated Fas-stimulated migration. We conclude that alpha(v)beta(8) integrin induction defines an unexpected role for Fas in cell migration, rather than as a cell death receptor.     

Application of dual colour FISH for localisation of canine microsatellite markers in the Chinese raccoon dog (Nyctereutes procyonoides procyonoides) genome

The present study shows new chromosomal localisation of four canine cosmid clones in the Chinese raccoon dog genome by using dual colour FISH. This approach facilitates rapid physical localisation of markers and will improve the determination of their order on chromosomes. The present new assignments increase the number of physically mapped markers in the Chinese raccoon dog to 25.     

Characterization and mapping of canine microsatellites isolated from BAC clones harbouring DNA sequences homologous to seven human genes

Human primers specific for the genes LEP, HBB, PAX3, ESR2, TPH1, ABCA4 and ATP2A2 were used to identify clones in a canine BAC library. Subcloning of the positive BACs in plasmids, screening with microsatellite motifs and subsequent sequencing allowed for the identification of eight novel microsatellites. The presence of the gene of interest was confirmed by sequencing the polymerase chain reaction (PCR) products amplified in the positive BACs. Fluorescent in situ hybridization (FISH) using the positive BACs as probes allowed for the chromosomal localization of the insert DNAs in two canid species, dog (Canis familiaris) and red fox (Vulpes vulpes). The use of gene-associated microsatellites may accelerate the identification of candidate genes for phenotypic traits in linkage studies.     

Na(+)/H(+) exchange inhibition prevents endothelial dysfunction after I/R injury

Whereas inhibition of the Na(+)/H(+) exchanger (NHE) has been demonstrated to reduce myocardial infarct size in response to ischemia-reperfusion injury, the ability of NHE inhibition to preserve endothelial cell function has not been examined. This study examined whether NHE inhibition could preserve endothelial cell function after 90 min of regional ischemia and 180 min of reperfusion and compared this inhibition with ischemic preconditioning (IPC). In a canine model either IPC, produced by one 5-min coronary artery occlusion (1 x 5'), or the specific NHE-1 inhibitor eniporide (EMD-96785, 3.0 mg/kg) was administered 15 min before a 90-min coronary artery occlusion followed by 3 h of reperfusion. Infarct size (IS) was determined by 2,3,5-triphenyl tetrazolium chloride staining and expressed as a percentage of the area-at-risk (IS/AAR). Endothelial cell function was assessed by measurement of coronary blood flow in response to intracoronary acetylcholine infusion at the end of reperfusion. Whereas neither control nor IPC-treated animals exhibited a significant reduction in IS/AAR or preservation of endothelial cell function, animals treated with the NHE inhibitor eniporide showed a marked reduction in IS/AAR and a significantly preserved endothelial cell function (P < 0.05). Thus NHE-1 inhibition is more efficacious than IPC at reducing IS/AAR and at preserving endothelial cell function in dogs.     

Immunochemical comparison of peptide angiogenic factors

A panel of 40 monoclonal antibodies was constructed in response to cationic endothelial cell growth factor (c-ECGF), the cationic peptide mitogen isolated from endothelial mitogen. The monoclonal antibodies were assayed by dot blot for immunoreactivity to various other peptide angiogenic factors. The panel of monoclonal antibodies to c-ECGF exhibited complete cross-reactivity with pituitary fibroblast growth factor and sarcoma-derived growth factor. A group of 28 monoclonal antibodies was found to exhibit reactivity to anionic endothelial mitogen (a-ECGF), brain fibroblast growth factor, endothelial cell growth factor, and retina-derived growth factor. None of the monoclonal antibodies was found to react with epidermal growth factor or platelet-derived growth factor. These data provide an immunological basis for grouping heparin-binding endothelial cell growth factors into anionic and cationic groups.     

Representative Seroprevalences of Human and Livestock Brucellosis in Two Mongolian Provinces

Mongolia implemented a brucellosis livestock mass vaccination campaign from 2000 to 2009. However, the number of human cases did not decline since 2004 and the current epidemiological situation in Mongolia was uncertain. The objective of this study was to estimate the representative seroprevalences of humans and livestock in two provinces in view of their comparison with officially reported data. A representative cross-sectional study using cluster sampling proportional to size in humans, sheep, goats, cattle, yaks, horses, camels and dogs was undertaken to assess the apparent seroprevalence in humans and animals. A total of 8054 livestock and dog sera and 574 human sera were collected in Sukhbaatar and Zavkhan provinces. Human and animal sera were tested with the Rose Bengal and ELISA tests. The overall apparent seroprevalence of brucellosis was 27.3% in humans (95% CI 23.7–31.2%), 6.2% (95% CI 5.5–7.1%) in sheep, 5.2% (95% CI 4.4–5.9%) in goats, 16.0% (95% CI 13.7–18.7%) in cattle, 2.5% (95% CI 0.8–7.6%) in camels, 8.3 (95% CI 6.0–11.6%) in horses and 36.4% (95% CI 26.3–48.0%) in dogs. More women than men were seropositive (OR = 1.7; P < 0.0014). Human seroprevalence was not associated with small ruminant and cattle seroprevalence at the nomadic camp (hot ail) level. Annual incidence of clinical brucellosis, inferred from the seroprevalence using a catalytic model, was by a factor of 4.6 (1307/280) in Sukhbaatar and by a factor of 59 (1188/20) in Zavkhan. This represents a 15-fold underreporting of human brucellosis in Mongolia. The lack of access to brucellosis diagnostic testing at the village level hinders rural people from receiving appropriate treatment. In conclusion, this study confirms the high seroprevalence of human and livestock brucellosis in Mongolia. Stringent monitoring and quality control of operational management of a nationwide mass vaccination of small and large ruminants is warranted to assure its effectiveness. More research is needed to understand the complex animal–human interface of brucellosis transmission at different scales from farm to provincial level.